Eisenia Fetida Research Articles

Earthworm (Oligochaeta) diversity of Kumaun Himalaya with a new record of Drawida japonica (Michaelsen, 1892) (Monaligastridae) from Nainital, Uttarakhand, India

The present study was conducted to investigate earthworm diversity in the Kumaun region of Uttarakhand, India. Earthworm sampling was made from 1,409–2,224 m from the foothills of the Kumaun Himalaya to higher altitudes monthly from April 2019 to April 2021, covering the major monsoon season in India. Earthworms were collected from twelve different sites using the hand sorting method and preserved in 10% formalin. We recorded 14 earthworm species representing four families: Megascolecidae (Amynthas corticis, Amynthas morrisi, Amynthas alexandri, Metaphire anomala, Metaphire houlleti, Metaphire birmanica, & Perionyx excavatus); Lumbricidae (Aporrectodea caliginosa trapezoides, Aporrectodea rosea rosea, Eisenia fetida, Octolasion tyrtaeum, & Dendrodrilus rubidus); Moniligastridae (Drawida japonica); and Octochaetidae (Eutyphoeus nainianus). Drawida japonica from the family Moniligastridae is being reported for the first time from the Kumaun Himalaya. Moniligastrids are an important, yet often ignored, earthworm group commonly found in cultivated soils of higher altitudes.

Toxicokinetics of selenate in earthworm sub-tissues and potential bio-accessibility assessment of earthworm-derived selenium

Selenium (Se) pollution is mainly caused by anthropogenic activities, and the resulting biosecurity concerns have garnered significant attention in recent years. Using one-compartmental toxicokinetic (TK) modelling, this study explored the kinetic absorption, sub-tissue distribution, and elimination processes of the main Se species (selenate, Se(VI)) in the cultivated aerobic soil of the earthworm Eisenia fetida. The bio-accessibility of earthworm-derived Se was assessed using an in vitro simulated gastrointestinal digestion test to evaluate its potential trophic risk. The results demonstrated that Se accumulated in the pre-clitellum (PC) and total tissues (TT) of earthworms in a time- and dose-dependent manner. The highest Se levels in the PC, post-clitellum (PoC), and TT were 70.54, 57.93, and 64.26 mg/kg during the uptake phase, respectively. The kinetic Se contents in the earthworms PC and TT were consistent with the TK model but not with PoC. The earthworm TT exhibited a faster uptake (Kus = 0.83–1.02 mg/kg/day) and elimination rate of Se (Kee = 0.044–0.049 mg/kg/day), as well as a shorter half-life time (LT1/2 = 15.88–14.22 days) than PC at low soil Se levels (≤5 mg/kg). Conversely, the opposite trend was observed with higher Se concentrations (10 and 20 mg/kg). These results are likely attributable to the tissue specificity and concentration of the toxicant. Earthworms PC and TT exhibited a higher kinetic Se accumulation factor (BAFk) than steady-state BAF (BAFss), with values ranging from 8 to 24 and 3–13, respectively. Furthermore, the bio-accessibility of earthworm-derived Se to poultry ranged from 66.25 % to 84.35 %. As earthworms are at the bottom of the terrestrial food chain, the high bio-accessibility of earthworm-derived Se poses a potential risk to predators. This study offers data support and a theoretical foundation for understanding the biological footprint of soil Se and its toxicological impacts and ecological hazards.

Safety and efficacy indicators of guarana and Brazil nut extract carried in nanoparticles of coenzyme Q10: Evidence from human blood cells and red earthworm experimental model

This study characterized a nanosupplement based on coenzyme Q10 containing guarana (Paullinia cupana) and Brazil nuts oil (Bertholetia excelsa) (G-Nut). Determined cytotoxic and oxi-immunomodulatory effects on human peripheral blood mononuclear cells (PBMCs) and its effect on mortality of red Californian earthworms (Eisenia fetida) and on the immune efficiency of its coelomocytes immune by in vitro exposure to yeast dead microorganism. The cytotoxic and immunomodulatory effects of G-Nut and the GN-Free extract (0.25–3 mg/mL) were determined in PBMC cultures. Apoptotic, oxidative, and inflammatory markers were determined using biochemical, immunological, and molecular protocols. The effects of G-Nut and GN-Free extracts on mortality and immune efficiency were investigated in earthworms. G-Nut and GN-Free did not induce cytotoxic events in PBMCs, triggering the decrease in apoptotic (caspases 3 and 8) gene expression, lipid and protein oxidation levels, or pro-inflammatory cytokine levels. G-Nut and GN-Free did not trigger earthworm mortality and improved coelomocyte immune efficiency by increasing Eisenia neutrophil extracellular DNA traps and brown body formation when exposed to dead yeasts. The G-Nut nanoformulation is safe and can be used as a new form of food supplement by oral or transdermal delivery.

Earthworms and lactic acid bacteria (LAB) cooperate to promote the biodegradation of tetracycline residues in livestock manure

Tetracycline is an antibiotic with extensive veterinary use in the livestock industry. However, their widespread application poses risks to soil health as residue in livestock feces, and their removal is crucial for sustainable soil-ecosystem development. Physical and chemical approaches to extract tetracycline may have adverse effects on soil ecosystems, but no studies have thus far examined the potential for biological methods, such as collective degradation action of soil fauna. Thus, this study aimed to investigate the synergistic effects of lactic acid bacteria (LAB) and earthworms (Eisenia fetida) on biodegradation of tetracycline residues in sheep manure. We assessed earthworm biomass, tetracycline residue, and bacterial communities in both earthworm intestines and vermicompost. Earthworm biomass and tetracycline degradation efficiency increased significantly with LAB addition, with a degradation rate of up to 80.16%. This increase may be attributable to LAB acting as electron donors to spur tetracycline degradation. Additionally, we noted that tetracycline presence significantly influenced bacterial communities in earthworm intestines and vermicompost, elevating the abundance of potential pathogenic bacteria (e.g., Flavobacterium, Gammaproteobacteria, and Enterobacteriaceae). This finding suggests that heightened environmental stress from antibiotics could actually facilitate the growth of less prevalent bacteria, including potential pathogens. In conclusion, our study provides evidence supporting the effectiveness of LAB and earthworms in degrading tetracycline residues. In particular, LAB appears to mitigate stress from tetracycline exposure in earthworms, thus increasing their vermicomposting efficacy. Our work has important implications for soil management, with the potential to enhance pollution clean-up rates while minimizing negative side-effects to soil microbial communities.

Role of root exudates on the transformation and ecological effect of engineering nanomaterials in soil system: A critical review

AbstractDue to the widespread use of engineered nanomaterials (ENMs) for soil remediation and nano‐enabled sustainable agriculture, there is a growing concern regarding the behavior and fate of ENMs released into soil systems in the presence of natural root exudates (REs). Herein, we investigate the influence of REs on the fate and ecological effect of ENMs from a comprehensive perspective. We summarize the key roles reported in the literature for REs in physical changes (e.g., adsorption, dispersion/aggregation), chemical changes (e.g., oxidation/redox reactions, and dissolution), and biotransformation of ENMs, which will further determine the ecological risk of ENMs in natural soil systems. Moreover, this review highlights the potential adverse effects of ENMs on different soil organisms (e.g., bacteria, plants, and eisenia foetida) in the presence of REs. The remaining unclear mechanisms (e.g., oxidative stress and DNA damage) of ENMs toxicity at the cellular level influenced by REs are reviewed and presented. Finally, the review concludes by addressing the current knowledge gaps and challenges in this field.

Vermicomposting preferably alters fungal communities in wasted activated sludge and promotes the production of plant growth-promoting biostimulants in the vermicompost

Waste activated sludge (WAS) is the main by-product of wastewater treatment plants. Vermicomposting integrated earthworms and reconstructive microbes to achieve WAS stabilization, with the mature composts enriched with plant hormones and humic substances (HS) forming supramolecular forces to protect hormone activity. However, how vermicomposting changes sludge microbial community, transforms sludge-derived organics into various plant biostimulants and promotes hormones-HS interaction remain elusive. Here we used multi-omics approaches to demonstrate how reconstructed microbes transformed sludge-derived organics into plant biostimulants, and molecular dynamics (MD) simulation to explore the force details of hormone-HS aggregations. Additionally, plant growth phenotype treated by the vermicompost was accessed. Our results showed that earthworm (Eisenia fetida) selectively altered fungal communities in sludge matrices, promoting the growth of phyla Mucoromycota, Ascomycota, and Cryptomycota. Reconstructed fungal communities then dominated the transformation of nitrogenous organics, accelerating the degradation rate of proteins and the production rate of plant biostimulants, including humic substances (HS, especially fulvic acid), indoleacetic acid (IAA), jasmonic acid (JA), and salicylic acid (SA). In detail, indole acted as the exclusive precursor for IAA biosynthesis (2.77 mg/kg-TS) via tryptophan metabolism. Unsaturated fatty acids generated from the citrate cycle were responsible for JA production (2.19 mg/kg-TS) via β-oxidation, while phenylalanine contributed to SA production (4.52 μg/kg-TS) via deaminase coenzyme and phenylalanine-4-hydroxylase. The results of MD simulation exhibited a stronger supramolecular force in the IAA-HS assemblage (−46.7 KJ/mol) than the SA-HS pair and JA-HS pair, thus protecting the structure and bioavailability of IAA and promoting the auxin-like activity in the vermicompost. Plant growth experiments further confirmed that vermicompost promoted plant root elongation. This work is believed to deepen the understanding of vermicomposting for sustainable management of sludge.

Novel mechanistic insights into Cr(VI) and Cr(III) induced discrepancies of cellular toxicity and oxidative injury events in Eisenia fetida

Chromium (Cr) poses a high ecological risk, however the toxic mechanisms of Cr in different valence states to soil organisms at cellular and molecular levels are not exactly. In this study, the Eisenia fetida coelomocytes and Cu/Zn-superoxide dismutase (Cu/Zn-SOD) were chosen as the target subjects to investigate the effects and mechanisms of cellular toxicity induced by Cr(VI) and Cr(III). Results indicated that Cr(VI) and Cr(III) significantly reduced the coelomocytes viability. The level of reactive oxygen species (ROS) was markedly increased after Cr(VI) exposure, which finally reduced antioxidant defense abilities, and induced lipid peroxidation and cellular membrane damage in earthworm coelomocytes. However, Cr(III) induced lower levels of oxidative stress and cellular damage with respect to Cr(VI). From a molecular perspective, the binding of both Cr(VI) and Cr(III) with Cu/Zn-SOD resulted in protein backbone loosening and reduced β-Sheet content. The Cu/Zn-SOD showed fluorescence enhancement with Cr(III), whereas Cr(VI) had no obvious effect. The activity of Cu/Zn-SOD continued to decrease with the exposure of Cr. Molecular docking indicated that Cr(III) interacted more readily with the active center of Cu/Zn-SOD. Our results illustrate that oxidative stress induced by Cr(VI) and Cr(III) plays an important role in the cytotoxic differences of Eisenia fetida coelomocytes and the binding of Cr with Cu/Zn-SOD can also affect the normal structures and functions of antioxidant defense-associated protein.

Comparative Study of Vermicomposting: Apple Pomace Alone and in Combination with Wheat Straw and Manure

Considering the sporadic number of scientific studies on vermicomposting apple pomace waste, this research conducts a comparative analysis of vermicomposting processes using Eisenia fetida, focusing on apple pomace both independently and in combination with wheat straw and/or manure (experiment 1: 60% apple pomace and 40% cattle manure; experiment 2: 60% wheat straw and 40% cattle manure; experiment 3: 80% apple pomace, 10% wheat straw, and 10% cattle manure; and experiment 4: 100% apple pomace). After a 240-day substrate transformation period, all four variations of vermicompost produced demonstrated favorable sensory properties, along with high microbiological and physicochemical quality. Throughout the vermicomposting process, the pH of all vermicomposting mixtures changed, converging towards approximately neutral values by the process’s conclusion. There was an increase in dry matter content, as well as total N, P, K, Ca, and Mg, along with organic matter. Notably, the levels of heavy metals (Zn, Cu, Cd, and Pb) in both the vermicomposting materials and resulting vermicomposts remained significantly below the maximum permissible levels stipulated by Republic of Serbia and European Union legislation, which is directly linked to the ecological origin of the raw materials used. The microbiological quality of the final vermicomposts was deemed satisfactory. Over time, there was a decrease in the counts of aerobic mesophilic bacteria as well as Escherichia coli. The counts of sulfite-reducing clostridia in all substrates remained below 102 CFU/g, while Salmonella spp. and Listeria monocytogenes were not detected in either the composting materials or the resulting composts. The vermiculture of apple pulp exhibited advantageous characteristics, notably a shortened vermicomposting period (150 days) compared to other agricultural waste. This reduction in processing time contributes an additional layer of advantage to the overall quality and efficiency of the resulting vermicompost.

A Bibliometric Analysis on Research Progress of Earthworms in Soil Ecosystems.

The earthworm, as a soil engineer, plays highly important roles in the soil ecosystem for shaping soil structure, promoting soil fertility, regulating microbial community composition and activities and decomposing soil pollutants. However, the research progresses on this important soil fauna have rarely been reviewed so far. Therefore, we conducted a bibliometric analysis of the literature published during 1900-2022, which was collected from the Web of Science Core Collection (WoS). The results showed that three periods (1900-1990, 1991-2005 and 2006-2022) could be identified in terms of the intensity of publications on the topic, and the number of publications kept increasing since 2006. The United States produced the highest publication record at the country scale, whereas Chinese Academy of Sciences was the most productive institution. Chinese institutions and authors played an active and prominent role during 2018-2022. Soil Biology & Biochemistry was the most popular journal for the topic-related research. In these publications, Professor Lavelle P was the most influential author. Based on a citation network of the top 50 cited papers, four hotspots were identified, i.e., the ecological effects of earthworms, the impact of agricultural activities on earthworms, earthworm ecotoxicology and earthworm invasion. Moreover, "impact", "biodiversity", "oxidative stress", "diversity", "response", "Eisenia fetida" and "exposure" were the emerging and active topics in recent years. This study can help us to better understand the relevant subject categories, journals, countries, institutions, authors and articles and identify the research hotspots and emerging trends in the field of soil earthworm research.

Use of biochar to improve sewage sludge quality in Maluti-A-Phofung Municipality, South Africa

Research on wastewater treatment processes in Maluti-a-Phofung Municipality (South Africa) has revealed that substandard wastewater management in this region contributes to terrestrial and aquatic pollution. Because this pollution poses a threat to the environment, there is a pressing need to reduce the environmental impact of poorly managed sewage sludge in the region. Biochar has been regarded as a cost-effective way of reducing chemical toxicity in terrestrial environments. In the present study, we aimed to investigate the effectiveness of biochar in the remediation of the toxicity of sewage sludge using the earthworm Eisenia fetida. Sewage sludge was collected from a local wastewater treatment plant and E. fetida were exposed to 0, 25, 50, and 100% non-amended and 10% biochar-amended sludge. After 28 days, survival, biomass and reproduction were assessed. Separately, in clean artificial soil, E. fetida was exposed to 5, 10 and 15% biochar amendment for 96 hours to determine if biochar amendment alone could be harmful to E. fetida. The results showed no significant differences in all parameters between the worms exposed to non-amended sludge and 10% biochar-amended sludge. Assessment of acetylcholinesterase and catalase activities in the earthworms that were exposed to biochar via clean soil revealed that 10% and 15% biochar amendment rates caused the worms to experience significant levels of neurotoxic and oxidative stress (p < 0.05). These findings reveal that biochar alone is likely to have adverse effects on soil organisms, and amendment rates higher or equal to 10% are not suitable to alleviate the toxic effects of sewage sludge.

Toxicity assessment of nano-hydroxyapatite, -zeolite, and -iron tetroxide to soil fauna: The earthworm (Eisenia fetida) as a model organism

The toxicity of nanoparticles has attracted much attention, but the toxicity of nanoparticle types to soil fauna remains unclear. In this study, the earthworm (Eisenia fetida) was selected as a representative soil fauna to evaluate the toxicity of nano-hydroxyapatite (n-HAP), -zeolite (n-zeolite), and -iron tetroxide (n-Fe3O4) to soil fauna. The results showed that the skin ultrastructure, mortality, growth, antioxidant systems, and genetic material of the earthworms were affected to varying degrees by different nanoparticles. A filter paper test indicated that dermal exposure to nanoparticles primarily caused lesions in the cuticle, microvilli, and nucleus of the epithelial cells of earthworm skin. Soil incubation experiments showed that all types of nanoparticles could cause weight loss in earthworms, but only n-Fe3O4 resulted in their death. The reactive oxygen species and glutathione contents in earthworms exposed to n-Fe3O4 were much higher than those in earthworms exposed to n-HAP and n-zeolite. All nanoparticles stimulated the catalase and lactate dehydrogenase activity and damaged the DNA of earthworms. Integrated biomarker responses showed that the toxicity to earthworms increased in the following order: n-HAP < n-zeolite < n-Fe3O4. The characterization results indicated that the differences in surface hydroxyl groups and surface charge may be the cause of the differences in toxicity between n-HAP and n-zeolite with similar properties. The highest toxicity of n-Fe3O4 could be due to the size, spherical morphology, positive surface charge and the presence of Fe(II).

Assessment of the Hazard and Ecotoxicity of the Residues of Paiche Leather Dyed with Natural and Synthetic Colourants: A Focus on Environmental Sustainability

The aim of this research was to assess the hazardousness and ecotoxicity of paiche leather (Arapaima gigas) dyed with natural colourants (turmeric and purple corn) versus synthetic colourants (anilines). To determine hazardousness, the methodology included the assessment of flammability, corrosivity, reactivity to H2S and HCN, toxicity from metals, and organic compounds of dyed leather. Additionally, its ecotoxicity was determined using the California red worm (Eisenia fetida) as a bioindicator through a bioassay, with treatments at 0.02, 0.04, 0.08, 0.16, and 0.20 g of leather/g of substrate, including undyed leather and a negative control in the investigation. The hazardousness analysis results indicated that leather samples dyed with both natural and synthetic colourants are not hazardous. However, the concentrations of chromium, nickel, cadmium, and lead exceed the permitted values for their use in footwear according to the European Union. Regarding ecotoxicity, the results demonstrate that acute toxicity to E. fetida is lower for natural colourants compared to synthetic colourants, where 100% mortality occurs at concentrations between 0.04 and 0.08 g of leather/g of the substrate. Concerning chronic toxicity, after 12 weeks of exposure, the tolerance concentration for undyed paiche leather is 0.035 g of leather/g of the substrate, and for leather dyed with purple corn and turmeric, it is 0.2 g of leather/g of the substrate. It is shown that the active compounds of natural colourants have a positive effect on worm survival, and it is concluded that paiche leather dyed with these colourants presents environmental advantages over leather dyed with synthetic colourants.

Combined effects of cadmium and sulfamethoxazole on Eisenia fetida: Insights into accumulation, subcellular partitioning, biomarkers and toxicological responses

Cadmium (Cd) and sulfamethoxazole (SMX) frequently coexist in farmlands, yet their synergistic toxicological impacts on terrestrial invertebrates remain unexplored. In this study, earthworms were exposed to artificial soils percolated with Cd (5 mg/kg), SMX (5 mg/kg) or combination of them for 7 days, followed by a 12-day elimination phase in uncontaminated soil. The uptake of Cd and SMX by the earthworms, along with their subcellular distribution, was meticulously analyzed. Additionally, a suite of biomarkers—including superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA), and weight loss—were evaluated to assess the health status of the earthworms and the toxicological effects of the Cd and SMX mixture. Notably, the cotreatment with Cd and SMX resulted in a significantly higher weight loss in Eisenia fetida (41.25 %) compared to exposure to Cd alone (26.84 %). Moreover, the cotreatment group exhibited substantially higher concentrations of Cd in the total internal body, fraction C (cytosol), and fraction E (tissue fragments and cell membranes) in Eisenia fetida compared to Cd alone counterparts. The combined exposure also significantly elevated the SMX levels in the total body and fraction C compared with the SMX-only treated earthworms. Additionally, Eisenia fetida subjected to the combined treatment showed markedly increased activities of SOD, CAT, and MDA compared to those treated with Cd alone. The effect addition indices (EAIs), ranging from 1.00 to 2.23, unequivocally demonstrated a synergistic effect of the combined treatments. Interestingly, relocating the earthworms to clean soil did not mitigate the observed adverse effects. These findings underscore the increased risk posed by the Cd-SMX complex to terrestrial invertebrates in agricultural areas.

Earthworms improve the rhizosphere micro-environment to mitigate the toxicity of microplastics to tomato (Solanum lycopersicum)

Microplastics (MPs) are widespread in agricultural soil, potentially threatening soil environmental quality and plant growth. However, toxicological research on MPs has mainly been limited to individual components (such as plants, microbes, and animals), without considering their interactions. Here, we examined earthworm-mediated effects on tomato growth and the rhizosphere micro-environment under MPs contamination. Earthworms (Eisenia fetida) mitigated the growth-inhibiting effect of MPs on tomato plant. Particularly, when exposed to environmentally relevant concentrations (ERC, 0.02% w/w) of MPs, the addition of earthworms significantly (p < 0.05) increased shoot and root dry weight by 12–13% and 13–14%, respectively. MPs significantly reduced (p < 0.05) soil ammonium (NH4+-N) (0.55–0.69 mg/kg), nitrate nitrogen (NO3−-N) (7.02–8.65 mg/kg) contents, and N cycle related enzyme activities (33.47–42.39 μg/h/g) by 37.7–50.9%, 22.6–37.2%, and 34.2–48.0%, respectively, while earthworms significantly enhanced (p < 0.05) inorganic N mineralization and bioavailability. Furthermore, earthworms increased bacterial network complexity, thereby enhancing the robustness of the bacterial system to resist soil MPs stress. Meanwhile, partial least squares modelling showed that earthworms significantly influenced (p < 0.01) soil nutrients, which in turn significantly affected (p < 0.01) plant growth. Therefore, the comprehensive consideration of soil ecological composition is important for assessing MPs ecological risk.

Impact of earthworms on antibiotic resistance genes removal in ampicillin-contaminated soil through bacterial community alteration.

The emergence of antibiotic resistance genes (ARGs) as contaminants in soil poses a significant threat to public health. Earthworms (Eisenia foetida), which are common inhabitants of soil, have been extensively studied for their influence on ARGs. However, the specific impact of earthworms on penicillin-related ARGs remains unclear. In this study, we investigate the role of earthworms in mitigating ARGs, specifically penicillin-related ARGs, in ampicillin-contaminated soil. Utilizing high-throughput quantitative PCR (HT-qPCR), we quantified a significant reduction in the relative abundance of penicillin-related ARGs in soil treated with earthworms, showing a decrease with a p-value of <0.01. Furthermore, high-throughput 16S rRNA gene sequencing revealed that earthworm intervention markedly alters the microbial community structure, notably enhancing the prevalence of specific bacterial phyla such as Proteobacteria, Firmicutes, Chloroflexi, and Tenericutes. Our findings not only demonstrate the effectiveness of earthworms in reducing the environmental load of penicillin-related ARGs but also provide insight into the alteration of microbial communities as a potential mechanism. This research contributes to our understanding of the role of earthworms in mitigating the spread of antibiotic resistance and provides valuable insights for the development of strategies to combat this global health issue.

Toxicity evaluation of neonicotinoids to earthworm (Eisenia fetida) behaviors by a novel locomotion tracking assay

Pesticides are substances used for controlling, preventing, and repelling pests in agriculture. Among them, neonicotinoids have become the fastest-growing class of insecticides because of their efficiency in targeting pests. They work by strongly binding to nicotinic acetylcholine receptors (nAChRs) in the central nervous system of insects, leading to receptor blockage, paralysis, and death. Despite their selectivity for insects, these substances may be hazardous to non-target creatures, including earthworms. Although earthworms may be invasive in some regions like north America, they contribute to the development of soil structure, water management, nutrient cycling, pollution remediation, and cultural services, positively impacting the environment, particularly in the soil ecosystem. Thus, this study aimed to develop a novel earthworm behavior assay since behavior is a sensitive marker for toxicity assay, and demonstrated its application in evaluating the toxicity of various neonicotinoids. Here, we exposed Eisenia fetida to 1 and 10 ppb of eight neonicotinoids (acetamiprid, clothianidin, dinotefuran, imidacloprid, nitenpyram pestanal, thiacloprid, thiametoxam, and sulfoxaflor) for 3 days to observe their behavior toxicities. Overall, all of the neonicotinoids decreased their locomotion, showed by a reduction of average speed by 24.94–68.63% and increment in freezing time movement ratio by 1.51–4.25 times, and altered their movement orientation and complexity, indicated by the decrement in the fractal dimension value by 24–70%. Moreover, some of the neonicotinoids, which were acetamiprid, dinotefuran, imidacloprid, nitenpyram, and sulfoxaflor, could even alter their exploratory behaviors, which was shown by the increment in the time spent in the center area value by 6.94–12.99 times. Furthermore, based on the PCA and heatmap clustering results, thiametoxam was found as the neonicotinoid that possessed the least pronounced behavior toxicity effects among the tested pesticides since these neonicotinoid-treated groups in both concentrations were grouped in the same major cluster with the control group. Finally, molecular docking was also conducted to examine neonicotinoids' possible binding mechanism to Acetylcholine Binding Protein (AChBP), which is responsible for neurotransmission. The molecular docking result confirmed that each of the neonicotinoids has a relatively high binding energy with AChBP, with the lowest binding energy was possessed by thiametoxam, which consistent with its relatively low behavior toxicities. Thus, these molecular docking results might hint at the possible mechanism behind the observed behavior alterations. To sum up, the present study demonstrated that all of the neonicotinoids altered the earthworm behaviors which might be due to their ability to bind with some specific neurotransmitters and the current findings give insights into the toxicities of neonicotinoids to the environment, especially animals in a soil ecosystem.

Joint toxic mechanism of clothianidin and prochloraz in the earthworm (Eisenia fetida)

Pesticides are typically present as combinations within soil ecosystems and have detrimental effects on untamed surroundings. However, the collective impacts and fundamental mechanisms of pesticides on soil living beings are currently inadequately assessed. In our current work, we evaluated the interactive consequences of clothianidin (CLO) and prochloraz (PRO) on earthworms (Eisenia fetida) using several toxicological tests, such as acute adverse effects, biocatalytic activity, and alterations in transcriptional activity. The findings revealed that CLO (with a 14-day LC50 value of 6.08 mg kg−1) exhibited greater toxicity compared to PRO (with a 14-day LC50 value of 79.41 mg kg−1). Moreover, the combinations of CLO and PRO had synergistic acute effects on E. fetida. Additionally, the activities of POD, CAT, and GST were significantly varied in most instances of single and mixed treatments when compared to the control. Surprisingly, the transcriptional levels of four genes (gst, sod, crt, and ann), related to oxidative load, metabolic detoxification systems, endoplasmic reticulum, and oxytocin neuropeptide, respectively, were also altered in response to single and mixture exposures, as compared to the control. Alterations in enzyme activity and gene transcriptional level could serve as early indicators for detecting co-exposure to pesticides. The findings of this research offered valuable holistic understanding regarding the toxicity of pesticide combinations on earthworms. Further research should be conducted to investigate the persistent effects of pesticide mixtures on terrestrial invertebrates in order to draw definitive conclusions about the associated risks.

Oxidative stress and DNA alteration on the earthworm Eisenia fetida exposed to four commercial pesticides

Modern agriculture is mainly based on the use of pesticides to protect crops but their efficiency is very low, in fact, most of them reach water or soil ecosystems causing pollution and health hazards to non-target organisms. Fungicide triazoles and strobilurins based are the most widely used and require a specific effort to investigate toxicological effects on non-target species. This study evaluates the toxic effects of four commercial fungicides Prosaro® (tebuconazole and prothioconazole), Amistar®Xtra (azoxystrobin and cyproconazole), Mirador® (azoxystrobin) and Icarus® (Tebuconazole) on Eisenia fetida using several biomarkers: lipid peroxidation (LPO), catalase activity (CAT), glutathione S-transferase (GST), total glutathione (GSHt), DNA fragmentation (comet assay) and lysozyme activity tested for the first time in E. fetida. The exposure to Mirador® and AmistarXtra® caused an imbalance of ROS species, leading to the inhibition of the immune system. AmistarXtra® and Prosaro®, composed of two active ingredients, induced significant DNA alteration, indicating genotoxic effects. This study broadened our knowledge of the effects of pesticide product formulations on earthworms and showed the need for improvement in the evaluation of toxicological risk deriving from the changing of physicochemical and toxicological properties that occur when a commercial formulation contains more than one active ingredient and several unknown co-formulants.

Reproduction toxicity study with the synthetic amorphous silica SYLOID® AL-1 FP, HDK® N20, LUDOX® P T-40 F and SYLOID® MX 107 in the earthworm species Eisenia fetida

Ecotoxicology studies were performed in the earthworm Eisenia fetida with four different synthetic amorphous silica (SAS) (SYLOID® AL-1 FP, SYLOID® MX 107, LUDOX® P T-40 F, and HDK® N20) mixed into artificial soil to determine a NOEC/LOEC for effects on reproduction (56 days after application), mortality and biomass development (28 days after application) using a standardized artificial soil with 10% peat. The LC50 for test-item effects on adult mortality, and an EC10 and EC50 for reproduction were also determined. Furthermore, earthworms underwent histopathology evaluation, and the amount of silica in different organs from these organisms was evaluated using EDX (Energy Dispersive X-ray Spectroscopy). Histopathology revealed no findings in any organ of the earthworms, except for desiccated dissepiments in evaluated decedents at extremely high SAS doses. To measure SAS uptake into the organs, a fully quantitative method for silica was established and validated using standards containing known concentrations of silica to ensure the accuracy of the analyses undertaken. Results from EDX analysis demonstrated the negligible presence of silicon within the brain ganglia and gonads of adult earthworms comparable to controls. Therefore, any deposition of the test items within these two organs was excluded. In contrast, traces of silicon higher than in controls were found in the intestinal lumina of the earthworms due to ingestion of SAS with soil and feed, but not in other organs.

Effects of Spirulina Platensis on Reproductive Rates of Earthworm (Eisenia Fetida) in Vermicomposting.

The purpose of the present study was to determine how adding Spirulina platensis to the diet would affect the growth and reproduction of Earthworm Eisenia fetida, in the process of vermicomposting. In the current study, vermicompost was prepared using cow dung. Total three setup were prepared for the both the treatment and control vermicom-posting. Development and reproduction of Eisenia fetida in cow dung feed supplemented with Spirulina platensis (in powdered form) were monitored weekly by counting the number of hatchlings and number of cocoons. When the vermicompost gets matured, cocoons, hatchlings and earthworm were removed to calculate the total change in biomass of Earthworm Eisenia fetida after the treatment. It was observed that Eisenia fetida fed with Spirulina platensis in vermicomposting had higher rates of reproduction than that of controlled vermicomposting. After the second week of treatment, premature worms formed clitella, and after the third week, cocoons were seen.